Anthranilic Acid Derivatives Active as the Hm74a Receptor

ABSTRACT

Therapeutically active anthranilic acid derivatives of Formula (I) or of Formula (Ia) wherein R, R 1 , R 2  and Z are as defined in the specification, processes for the preparation of said derivatives, pharmaceutical formulations containing the active compounds and the use of the compounds in therapy, particularly in the treatment of diseases in which under-activation of the HM74A receptor contributes to the disease or in which activation of the receptor will be beneficial, are disclosed.

The present invention relates to therapeutically active compounds whichare anthranilic acid derivatives, processes for the manufacture of saidderivatives, pharmaceutical formulations containing the active compoundsand the use of the compounds in therapy, particularly in the treatmentof diseases in which under-activation of the HM74A receptor contributesto the disease or in which activation of the receptor will bebeneficial.

Dyslipidaemia is a general term used to describe individuals withaberrant lipoprotein profiles. Clinically, the main classes of compoundsused for the treatment of patients with dyslipidaemia, and therefore atrisk of cardiovascular disease are the statins, fibrates, bile-acidbinding resins and nicotinic acid. Nicotinic acid (Niacin, a B vitamin)has been used clinically for over 40 years in patients with variousforms of dyslipidaemia. The primary mode of action of nicotinic acid isvia inhibition of hormone-sensitive triglyceride lipase (HSL), whichresults in a lowering of plasma non-esterified fatty acids (NEFA) whichin turn alters hepatic fat metabolism to reduce the output of LDL andVLDL (low and very low density lipoprotein). Reduced VLDL levels arethought to lower cholesterol ester transfer protein (CETP) activity toresult in increased HDL (high density lipoprotein) levels which may bethe cause of the observed cardiovascular benefits. Thus, nicotinic acidproduces a very desirable alteration in lipoprotein profiles; reducinglevels of VLDL and LDL whilst increasing HDL. Nicotinic acid has alsobeen demonstrated to have disease modifying benefits, reducing theprogression and increasing the regression of atherosclerotic lesions andreducing the number of cardiovascular events in several trials.

The observed inhibition of HSL by nicotinic acid treatment is mediatedby a decrease in cellular cyclic adenosine monophosphate (cAMP) causedby the G-protein-mediated inhibition of adenylyl cyclase. Recently, theG-protein coupled receptors HM74 and HM74A have been identified asreceptors for nicotinic acid (PCT patent application WO02/84298; Wiseet. al. J Biol Chem. 2003 278 (11) 9869-9874). The DNA sequence of humanHM74A may be found in Genbank; accession number AY148884. Two otherpapers support this discovery, (Tunaru et. al. Nature Medicine 2003 (3)352-255 and Soga et. al. Biochem Biophys Res Commun. 2003 303 (1)364-369), however the nomenclature differs slightly. In the Tunaru paperwhat they term human HM74 is in fact HM74A and in the Soga paper HM74bis identical to HM74A. Cells transfected to express HM74A and/or HM74gain the ability to elicit G_(i) G-protein mediated responses followingexposure to nicotinic acid. In mice lacking the homologue of HM74A(m-PUMA-G) nicotinic acid fails to reduce plasma NEFA levels.

We now present a group of anthranilic acid derivatives which areselective agonists of the nicotinic acid receptor HM74A and are thus ofbenefit in the treatment, prophylaxis and suppression of diseases whereunder-activation of this receptor either contributes to the disease orwhere activation of the receptor will be beneficial.

SUMMARY OF THE INVENTION

The present invention provides therapeutically active anthranilic acidderivatives and the use of these derivatives in therapy, particularly inthe treatment of diseases in which under-activation of the HM74Areceptor contributes to the disease or in which activation of thereceptor will be beneficial, in particular diseases of lipid metabolismincluding dislipidaemia or hyperlipoproteinaemia such as diabeticdyslipidaemia and mixed dyslipidaemia, heart failure,hypercholesterolaemia, cardiovascular disease including atherosclerosis,arteriosclerosis, and hypertriglyceridaemia. As such, the compounds mayalso find favour as therapeutics for coronary artery disease,thrombosis, angina, chronic renal failure, peripheral vascular diseaseand stroke, as well as the cardiovascular indications associated withtype II diabetes mellitus, type I diabetes, insulin resistance,hyperlipidaemia, anorexia nervosa, obesity. The compounds may also be ofuse in the treatment of inflammatory diseases or conditions, as set outfurther below.

Intermediates, formulations, methods and processes described herein formfurther aspects of the invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a compound of Formula (I)

and salts, solvates and physiologically functional derivatives thereof,wherein:R represents (CH₂)_(q)CO₂H or a 5 or 6-member aryl, heteroaryl,heterocyclic or alicyclic ring;R¹ represents H, halogen, C₁₋₅alkyl, C₁-C₅alkoxy, C₁₋₅haloalkyl, orC₁₋₅haloakloxy;R² represents a 5 or 6-member aryl, heteroaryl, heterocyclic oralicyclic ring;Z represents —(CH₂)_(q)—; —CH═CH—; —(CH₂)_(p)NHC(O)—;—(CH₂)_(p)NHC(O)NH—; —(CH₂)_(p)NHC(O)O—; —(CH₂)_(p)SO₂NR³—;—(CH₂)_(p)NR³SO₂—; —(CH₂)_(n)O—; —C(R⁴R⁵)O— or —Y—W—X—;W represents a 5 or 6-member aryl, heteroaryl, heterocyclic or alicyclicring;X and Y, which may independently be present or absent, where presentindependently represent —(CH₂)_(q)—; —CH═CH—; —(CH₂)_(p)NHC(O)—;—(CH₂)_(p)NHC(O)O—; —(CH₂)_(p)NHC(O)NH—; —(CH₂)_(p)SO₂NR³—;—(CH₂)_(p)NR³SO₂—; —(CH₂)_(p)C(O)—; —(CH₂)_(p)NH—; —(CH₂)_(p)O—;—(CH₂)_(p)S— or —(CH₂)_(p)O—CH₂—;n represents an integer selected from 2, 3 and 4;p represents an integer selected from 0, 1 and 2;q represents an integer selected from 1, 2, 3 and 4;R³ represents hydrogen or methyl; andR⁴ and R⁵, which may be the same or different, independently representC₁-C₃alkyl.

Also provided is a compound of Formula (Ia)

and salts, solvates and physiologically functional derivatives thereof,wherein:R represents CO₂H;R¹ represents C₁-C₅alkoxy, C₁₋₅haloalkyl, or C₁₋₅haloakloxy;Z and R² are as set out above.

In compounds of the present invention, the R² ring system may be joinedto the Z linker unit via either a ring carbon atom or via a ringheteroatom, where present. Similarly, where R represents a ring system,this may be joined to the phenyl moiety via either a ring carbon atom orvia a ring heteroatom, where present.

In certain compounds of Formula (I) in which R is heteroaryl, R may bepyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, imidazolyl,tetrazolyl, oxazolyl and isoxazolyl, for example tetrazolyl. Where Rrepresents (CH₂)_(q)CO₂H, in certain embodiments q represents 1 or 2,for example 1. In certain embodiments in which R is heterocyclic, R isselected from pyrrolidinyl, imidazolidinyl, piperidinyl and morpholinyl.

In further compounds of Formula (I), R is selected from cyclohexyl,phenyl, pyridinyl, pyrimidinyl, pyridazinyl, tetrazolyl and isoxazolyl.As defined below, the 5 or 6-member aryl, heteroaryl, heterocyclic oralicyclic R groups may be substituted and thus include substitutedcyclohexyl, substituted phenyl, substituted pyridine, substitutedpyrimidine, substituted pyridazine, substituted tetrazole or substitutedisoxazole, in which the substituents are as defined further below.

In certain compounds of Formula (I), R¹ groups are H, fluoro orC₁-C₃alkoxy, for example methoxy.

In certain compounds of Formula (Ia), R¹ groups are C₁-C₃alkoxy, forexample methoxy.

In certain embodiments of the present invention in which R² isheteroaryl, R² is selected from pyridinyl, pyrimidinyl, pyridazinyl,pyrazinyl, pyrazolyl, imidazolyl, oxazolyl and isoxazolyl. In certainembodiments in which R² is heterocyclic, R² is selected frompyrrolidinyl, imidazolidinyl, piperidinyl and morpholinyl.

In further embodiments, R² is selected from cyclohexyl, phenyl,pyridinyl, pyrimidinyl, pyridazinyl and isoxazolyl. As defined below,the 5 or 6-member aryl, heteroaryl, heterocyclic or alicyclic R² groupsmay be substituted and thus include substituted cyclohexyl, substitutedphenyl, substituted pyridine, substituted pyrimidine, substitutedpyridazine or substituted isoxazole, in which the substituents are asdefined further below.

Thus, where R² is substituted phenyl, the substituents will be thosedefined for “aryl” substituents below. In some embodiments thesubstituted phenyl bears one or two substituents selected from halogenC₁₋₃alkyl (for example methylphenyl), C₁₋₃haloalkyl (for exampletrifluoroalkyl including trifluoromethylphenyl), C₁₋₃alkoxy (for examplemethoxyphenyl) and C₁₋₃haloakloxy (for example trifluoroalkoxy includingtrifluoromethoxyphenyl).

In certain embodiments in which R² represents singly substituted phenyl,the substituent is at the meta or para position, for example para. Incertain embodiments in which R² represents doubly substituted phenyl,the substituents are at the para and meta, or at both meta positions.

In certain embodiments, R² is selected from the group consisting of:

In certain embodiments of the present invention, Z represents —Y—W—X—,—(CH₂)_(q)—, —(CH₂)_(n)O— or —(CH₂)_(p)NHC(O)—.

In certain embodiments Y represents —O—, —CH₂— or —CH₂O—. In particularembodiments, X is absent or represents —(CH₂)_(p)SO₂NR³—,—(CH₂)_(p)NHC(O)— or —(CH₂)_(p)NHC(O)NH—. In certain embodiments inwhich Y represents —CH₂—, X represents —(CH₂)_(p)SO₂NR³—. In certainembodiments in which Y represents —O— or —CH₂O—, X is absent.

Particular W groups are 5 or 6 member aryl or heteroaryl rings. Incertain embodiments in which W is aryl, for example C6 aryl (e.g.phenyl), W is linked through the 1 and 4 or the 1 and 3 positions. Incertain embodiments in which W is heteroaryl, for example a 5 memberheteroaryl ring (e.g. 1,2,4 oxadiazolyl), W may be linked through the 3and 5 positions. In other embodiments in which W is heteroaryl, forexample a 6 member heteroaryl ring (e.g. pyridinyl), W may be linkedthrough the 2 and 5 positions. When X is —(CH₂)_(p)SO₂NR³—, p is 0 and Wis unsubstituted phenyl, W may for example be linked through the 1 and 4(para) positions.

In certain embodiments, n represents 2.

In certain embodiments, p represents an integer selected from 0 or 1.

In certain embodiments W and R² each represent unsubstituted phenyl,whilst in other embodiments W represents unsubstituted phenyl and R²represents substituted phenyl.

It is to be understood that the present invention includes anycombination of particular embodiments and covers all combinations ofparticular substituents described hereinabove.

Throughout the present specification and the accompanying claims thewords “comprise” and “include” and variations such as “comprises”,“comprising”, “includes” and “including” are to be interpretedinclusively. That is, these words are intended to convey the possibleinclusion of other elements or integers not specifically recited, wherethe context allows

As used herein, the terms “halogen” or “halo” refer to fluorine,chlorine, bromine and iodine.

As used herein, the term “alkyl” (when used as a group or as part of agroup) refers to an optionally substituted straight or branchedhydrocarbon chain containing the specified number of carbon atoms. Forexample, C₁-C₃alkyl means a straight or branched hydrocarbon chaincontaining at least 1 and at most 3 carbon atoms.

Examples of alkyl as used herein include, but are not limited to; methyl(Me), ethyl (Et), n-propyl, i-propyl and the like. Unless otherwisestated, optional substituents include hydroxy, halogen, ═S and ═O.

As used herein, the term “alkoxy” (when used as a group or as part of agroup) refers to an alkyl ether radical, wherein the term “alkyl” isdefined above. Examples of alkoxy as used herein include, but are notlimited to; methoxy, ethoxy, n-propoxy, i-propoxy and the like.

As used herein, the term “alicyclic” (when used as a group or as part ofa group) refers to a cyclic hydrocarbon ring containing the specifiednumber of carbon atoms. Examples of alicyclic as used herein include,but are not limited to cyclohexyl, cyclopropyl and the like. Saidalicyclic groups may be optionally substituted with one or more, forexample 1 to 3, groups selected from hydroxy, halogen, ═S, ═O,C₁-C₃alkyl (which may be further substituted with one or more hydroxy,═O or halo groups), optionally halogenated C₁-C₃alkoxy,C₁-C₃alkoxyC₁-C₃alkyl, NR³ ₂, —NHC(O)C₁-C₃alkyl, —C(O)NR³ ₂, and—S(O)₂C₁-C₃alkyl, wherein R³ is as defined above.

As used herein, the term “aryl” (when used as a group or as part of agroup) refers to an aromatic hydrocarbon ring of the specified number ofcarbons. Examples of aryl as used herein include, but are not limitedto, phenyl and benzyl. Said aryl groups may be optionally substitutedwith one or more, for example 1 to 3 groups selected from hydroxy,halogen, ═S, ═O, C₁-C₃alkyl (which may be further substituted with oneor more hydroxy, ═O or halo groups), optionally halogenated C₁-C₃alkoxy,C₁-C₃alkoxyC₁-C₃alkyl, NR³ ₂, —NHC(O)C₁-C₃alkyl, —C(O)NR³ ₂, and—S(O)₂C₁-C₃alkyl, wherein R³ is as defined above.

As used herein, the term “heteroaryl” (when used as a group or as partof a group) refers to an aryl group, as defined above, which containsone or more nitrogen or oxygen heteroatoms. Examples of heteroaryl asused herein include, but are not limited to, pyridine, pyrimidine,pyridazine, imidazole, isoxazole, oxadiazoles, tetrazoles and the like.Said heteroaryl groups may be optionally substituted with one or more,for example 1 to 3 groups selected from hydroxy, halogen, ═S, ═O,C₁-C₃alkyl (which may be further substituted with one or more hydroxy,═O or halo groups), optionally halogenated C₁-C₃alkoxy,C₁-C₃alkoxyC₁-C₃alkyl, NR³ ₂, —NHC(O)C₁-C₃alkyl, —C(O)NR³ ₂, and—S(O)₂C₁-C₃alkyl, wherein R³ is as defined above.

As used herein, the term “heterocyclic” (when used as a group or as partof a group) refers to an alicyclic group, as defined above, whichcontains one or more nitrogen or oxygen heteroatoms. Said heterocyclicgroups may be optionally substituted with one or more, for example 1 to3 groups selected from hydroxy, halogen, ═S, ═O, C₁-C₃alkyl (which maybe further substituted with one or more hydroxy, ═O or halo groups),optionally halogenated C₁-C₃alkoxy, C₁-C₃alkoxyC₁-C₃alkyl, NR³ ₂,—NHC(O)C₁-C₃alkyl, —C(O)NR³ ₂, and —S(O)₂C₁-C₃alkyl, wherein R³ is asdefined above.

As used herein, the term “physiologically functional derivative” refersto any pharmaceutically acceptable derivative of a compound of thepresent invention, for example an ester or an amide thereof, andincludes any pharmaceutically acceptable salt, ester, or salt of suchester of a compound of Formula (I) or of Formula (Ia) which, uponadministration to a mammal, such as a human, is capable of providing(directly or indirectly) a compound of Formula (I) or of Formula (Ia) oran active metabolite or residue thereof. It will be appreciated by thoseskilled in the art that the compounds of Formula (I) and of Formula (Ia)may be modified to provide physiologically functional derivativesthereof at any of the functional groups in the compounds, and that thecompounds of Formula (I) and (Ia) may be so modified at more than oneposition.

As used herein, the term “pharmaceutically acceptable” used in relationto an ingredient (active ingredient or excipient) which may be includedin a pharmaceutical formulation for administration to a patient, refersto that ingredient being acceptable in the sense of being compatiblewith any other ingredients present in the pharmaceutical formulation andnot being deleterious to the recipient thereof.

As used herein, the term “solvate” refers to a complex of variablestoichiometry formed by a solute (in this invention, a compound ofFormula (I), a salt thereof or a physiologically functional derivativethereof, or a compound of Formula (Ia), a salt thereof or aphysiologically functional derivative thereof,) and a solvent. Suchsolvents for the purposes of the present invention may not interferewith the biological activity of the solute. Examples of suitablesolvents include water, methanol, ethanol and acetic acid. Preferablythe solvent used is a pharmaceutically acceptable solvent. Examples ofsuitable pharmaceutically acceptable solvents include water, ethanol andacetic acid. Most preferably the solvent used is water, in which casethe solvate may be referred to as a hydrate of the solute in question.

It will be appreciated that, for pharmaceutical use, the “salt orsolvate” referred to above will be a pharmaceutically acceptable salt orsolvate. However, other salts or solvates may find use, for example, inthe preparation of a compound of Formula (I) or of Formula (Ia) or inthe preparation of a pharmaceutically acceptable salt or solvatethereof.

Pharmaceutically acceptable salts include those described by Berge,Bighley and Monkhouse, J. Pharm. Sci., 1977, 66, 1-19. Suitablepharmaceutically acceptable salts include acid addition salts formedfrom the addition of inorganic acids or organic acids, preferablyinorganic acids. Examples of suitable acid addition salts includehydrochlorides, hydrobromides, sulphates and acetates. Furtherrepresentative examples of pharmaceutically acceptable salts includethose formed from maleic, fumaric, benzoic, ascorbic, pamoic, succinic,bismethylenesalicylic, methanesulfonic, ethanedisulfonic, propionic,tartaric, salicylic, citric, gluconic, aspartic, stearic, palmitic,itaconic, glycolic, p-aminobenzoic, glutamic, benzenesulfonic,cyclohexylsulfamic, phosphoric and nitric acids. Suitablepharmaceutically acceptable salts also include alkali metal salts formedfrom the addition of alkali metal bases such as alkali metal hydroxides.An example of a suitable alkali metal salt is a sodium salt.

In a further aspect, the present invention provides the use of acompound of Formula (I)

and salts, solvates and physiologically functional derivatives thereofas define above in the manufacture of a medicament for the treatment ofdisorders of lipid metabolism, including dislipidaemia orhyperlipoproteinaemia, or of inflammatory diseases or conditions. It isto be understood that this aspect of the present invention includes,with respect to the use of compounds of Formula (I) in the manufactureof a medicament, any combination of particular embodiments and coversall combinations of particular substituents of compounds of Formula (I)described hereinabove.

This aspect of the invention also provides the use of a compound ofFormula (Ia)

and salts, solvates and physiologically functional derivatives thereofas define above in the manufacture of a medicament for the treatment ofdisorders of lipid metabolism, including dislipidaemia orhyperlipoproteinaemia, or of inflammatory diseases or conditions. It isto be understood that this aspect of the present invention includes,with respect to the use of compounds of Formula (Ia) in the manufactureof a medicament, any combination of particular embodiments and coversall combinations of particular substituents of compounds of Formula (Ia)described hereinabove.

Compounds of the present invention are of potential therapeutic benefitin the treatment and amelioration of the symptoms of many diseases oflipid metabolism including dislipidaemia or hyperlipoproteinaemia suchas diabetic dyslipidaemia and mixed dyslipidaemia, heart failure,hypercholesterolaemia, cardiovascular disease including atherosclerosis,arteriosclerosis, and hypertriglyceridaemia. As such, the compounds mayalso find favour as therapeutics for coronary artery disease,thrombosis, angina, chronic renal failure, peripheral vascular diseaseand stroke, as well as the cardiovascular indications associated withtype II diabetes mellitus, type I diabetes, insulin resistance,hyperlipidaemia, anorexia nervosa, obesity. The use of a compound ofFormula (I) or a compound of Formula (Ia) in the treatment of one ormore of these diseases is a further aspect of the present invention.

Furthermore, it is also believed that the HM74 and HM74A receptors areinvolved in inflammation. Inflammation represents a group of vascular,cellular and neurological responses to trauma. Inflammation can becharacterised as the movement of inflammatory cells such as monocytes,neutrophils and granulocytes into the tissues. This is usuallyassociated with reduced endothelial barrier function and oedema into thetissues. Inflammation with regards to disease typically is referred toas chronic inflammation and can last up to a lifetime. Such chronicinflammation may manifest itself through disease symptoms. The aim ofanti-inflammatory therapy is therefore to reduce this chronicinflammation and allow for the physiological process of healing andtissue repair to progress.

Thus, a further aspect of the present invention resides in the use of acompound of Formula (I) or a salt, solvate or physiologically functionalderivative thereof or a compound of Formula (Ia) or a salt, solvate orphysiologically functional derivative thereof as defined above in thetreatment of inflammatory diseases or conditions of the joint,particularly arthritis (e.g. rheumatoid arthritis, osteoarthritis,prosthetic joint failure), or the gastrointestinal tract (e.g.ulcerative colitis, Crohn's disease, and other inflammatory bowel andgastrointestinal diseases, gastritis and mucosal inflammation resultingfrom infection, the enteropathy provoked by non-steroidalanti-inflammatory drugs), of the lung (e.g. adult respiratory distresssyndrome, asthma, cystic fibrosis, or chronic obstructive pulmonarydisease), of the heart (e.g. myocarditis), of nervous tissue (e.g.multiple sclerosis), of the pancreas, (e.g. inflammation associated withdiabetes melitus and complications thereof), of the kidney (e.g.glomerulonephritis), of the skin (e.g. dermatitis, psoriasis, eczema,urticaria, burn injury), of the eye (e.g. glaucoma) as well as oftransplanted organs (e.g. rejection) and multi-organ diseases (e.g.systemic lupus erythematosis, sepsis) and inflammatory sequelae of viralor bacterial infections and inflammatory conditions associated withatherosclerosis and following hypoxic or ischaemic insults (with orwithout reperfusion), for example in the brain or in ischaemic heartdisease.

In particular, the compounds of Formula (I) and (Ia) are useful in thetreatment and prevention of inflammation, and cardiovascular diseases orconditions including atherosclerosis, arteriosclerosis,hypertriglyceridemia, and mixed dyslipidaemia.

Thus, there is also provided the use of a compound of Formula (I) or apharmaceutically acceptable salt, solvate or physiologically functionalderivative thereof, or a compound of Formula (Ia) or a salt, solvate orphysiologically functional derivative thereof, in the manufacture of amedicament for the treatment of disorders of lipid metabolism includingdislipidaemia or hyperlipoproteinaemia such as diabetic dyslipidaemiaand mixed dyslipidaemia, heart failure, hypercholesterolaemia,cardiovascular disease including atherosclerosis, arteriosclerosis, andhypertriglyceridaemia. The compounds are also provided for use in thetreatment of coronary artery disease, thrombosis, angina, chronic renalfailure, peripheral vascular disease and stroke, as well as thecardiovascular indications associated with type II diabetes mellitus,type I diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa,obesity.

Nicotinic acid has a significant side effect profile, possibly becauseit is dosed at high level (gram quantities daily). The most common sideeffect is an intense cutaneous flushing. The compounds of the presentinvention preferably exhibit reduced side effects compared to nicotinicacid.

HM74A has been identified as a high affinity receptor for nicotinic acidwhilst HM74 is a lower affinity receptor. Desirably, the compounds ofthe present invention are selective for HM74A by which is meant thatthey show greater affinity for HM74A than for HM74.

The potential for compounds of Formula (I) and Formula (Ia) to activateHM74A may be demonstrated, for example, using the following in vitro andin vivo assays:

In-Vitro Testing

For transient transfections, HEK293T cells (HEK293 cells stablyexpressing the SV40 large T-antigen) are maintained in DMEM containing10% foetal calf serum and 2 mM glutamine. Cells are seeded in 90 mmculture dishes and grown to 60-80% confluence (18-24 h) prior totransfection. Human HM74A (GenBank™ accession number AY148884) issubcloned in to a mammalian expression vector (pcDNA3; Invitrogen) andtransfected using Lipofectamine reagent. For transfection, 9 μg of DNAis mixed with 30 μl Lipofectamine in 0.6 ml of Opti-MEM (LifeTechnologies Inc.) and incubated at room temperature for 30 min prior tothe addition of 1.6 ml of Opti-MEM. Cells are exposed to theLipofectamine/DNA mixture for 5 h and 6 ml of 20% (v/v) foetal calfserum in DMEM is then added. Cells are harvested 48 h aftertransfection. Pertussis toxin treatment is carried out bysupplementation into media at 50 ngml⁻¹ for 16 h. All transienttransfection studies involve co-transfection of receptor together withthe G_(i/o) G protein, G_(o1)α.

For generation of stable cell lines the above method is used totransfect CHO-K1 cells seeded in six well dishes grown to 30%confluence. These cells are maintained in DMEM F-12 HAM media containing10% foetal calf serum and 2 mM glutamine. 48 h post-transfection themedia is supplemented with 400 μg/ml Geneticin (G418, Gibco) forselection of antibiotic resistant cells. Clonal CHO-K1 cell lines stablyexpressing HM74A are confirmed by [³⁵S]-GTPγS binding measurements,following the addition of nicotinic acid.

P2 membrane preparation—Plasma membrane-containing P2 particulatefractions are prepared from cell pastes frozen at −80° C. after harvest.All procedures are carried out at 4° C. Cell pellets are resuspended in1 ml of 10 mM Tris-HCl and 0.1 mM EDTA, pH 7.5 (buffer A) and byhomogenisation for 20s with a Ultra Turrax followed by passage (5 times)through a 25-gauge needle. Cell lysates are centrifuged at 1,000 g for10 min in a microcentrifuge to pellet the nuclei and unbroken cells andP2 particulate fractions are recovered by microcentrifugation at 16,000g for 30 min. P2 particulate fractions are resuspended in buffer A andstored at −80° C. until required.

[³⁵S]-GTPγS binding—Assays are performed at room temperature either in96-well format as described previously (Wieland, T. and Jakobs, K. H.(1994) Methods Enzymol. 237, 3-13) or in an adapted protocol carried outin 384-well format.

96-well format: Briefly, membranes (10 μg per point) are diluted to0.083 mg/ml in assay buffer (20 mM HEPES, 100 mM NaCl, 10 mM MgCl₂,pH7.4) supplemented with saponin (10 mg/l) and pre-incubated with 10 μMGDP. Various concentrations of nicotinic acid or related molecules areadded, followed by [³⁵S]-GTPγS (1170 Ci/mmol, Amersham) at 0.3 nM (totalvol. of 100 μl) and binding is allowed to proceed at room temperaturefor 30 min. Non-specific binding is determined by the inclusion of 0.6mM GTP. Wheatgerm agglutinin SPA beads (Amersham) (0.5 mg) in 25 μlassay buffer are added and the whole is incubated at room temperaturefor 30 min with agitation. Plates are centrifuged at 1500 g for 5 minand bound [³⁵S]-GTPγS is determined by scintillation counting on aWallac 1450 microbeta Trilux scintillation counter.

384-well format: Briefly, the dilution of standard or test compounds areprepared and added to a 384-well plate in a volume of 10 μl. Membranes(HM74A or HM74) are diluted in assay buffer (20 mM HEPES, 100 mM NaCl,10 mM MgCl₂, pH7.4) supplemented with saponin (60 μg/ml), Leadseeker WGAbeads (Amersham; 250 μg/well) and 10 μM GDP, so that the 20 μl volumeadded to each well contains 5 μg of membranes. [³⁵S]-GTPγS (1170Ci/mmol, Amersham) is diluted (1:1500) in assay buffer and 20 μl addedto each well. Following the addition of the radioligand, the plates aresealed, pulse spun and incubated for 4 hours at room temperature. At theend of the incubation period the plates are read on a Leadseeker machine(VIEWLUX PLUS; Perkin-Elmer) to determine the levels of specificbinding.

In-Vivo Testing

HM74A agonists are tested in male Spague-Dawley rats (200-250 grams)which have been fasted for at least 12 hours prior to the study. Thecompounds are dosed intravenously (5 ml/kg) or by oral gavage (10ml/kg). Blood samples (0.3 ml tail vein bleed) are taken pre-dose and atthree times post-dose (times ranging from 15 minutes to 8 hourspost-dose). Each blood sample is transferred to a heparin tube (BectonDickinson Microtainer, PST LH) and centrifuged (10,000 g for 5 minutes)to produce a plasma sample. The plasma samples are assayed for levels ofnon-esterified fatty acids (NEFA) using a commercially available kit(Randox). Inhibition of plasma NEFA levels, relative to pre-dose levels,is used as a surrogate for HM74A agonist activity.

In order to determine whether compounds of the invention exhibit theflushing response associated with nicotinic acid, they are dosed toanaesthetised guinea-pigs. Nicotinic acid is used as positive control.Male Dunkin Hartley guinea pigs (300-800 g) are fasted for 12 hoursprior to being anaesthetised with a mixture of Ketamine hydrochloride(Vetalar, 40 mg/kg i.m.), Xylazine (Rompun, 8 mg/kg i.m.) and sodiumpentobarbitone (Sagatal, 30 mg/kg i.p.). Following anaesthesia atracheostomy is performed and the animals are mechanically ventilatedwith room air (10-12 mL/kg, 60 breaths/min). A jugular vein, and acarotid artery, are cannulated for intravenous administration of testcompound and collection of blood respectively. An infra-red temperatureprobe (Extech Instruments) is placed 3-5 mm from the tip of the leftear. Temperature measurements are recorded every minute from 5 minutesprior to test compound or nicotinic acid and up to 40 minutespost-administration of test compound or nicotinic acid. Data isautomatically collected on a Psion computer before being transferred fordata analysis within an Excel spreadsheet. Prior to, and at frequenttime points after compound administration, blood samples (0.3 ml) aretaken via the carotid arterial cannula and transferred to Microtainer(BD) tubes containing lithium heparin. The samples are mixed thoroughlyon a blood roller and then stored on ice prior to centrifugation at 1200g for 5 minutes.

Compounds according to Formulae (I) and (Ia) have been synthesised (seesynthetic examples below).

As indicated above, compounds of Formula (I) or of Formula (Ia) areuseful in human or veterinary medicine, in particular as activators ofHM74A, in the management of dyslipidaemia and hyperlipoproteinaemia.

Thus, there is provided as a further aspect of the present invention acompound of Formula (I) or a pharmaceutically acceptable salt, solvateor physiologically functional derivative thereof, or a compound ofFormula (Ia) or a pharmaceutically acceptable salt, solvate orphysiologically functional derivative thereof, for use in human orveterinary medicine, particularly in the treatment of disorders of lipidmetabolism including dislipidaemia or hyperlipoproteinaemia such asdiabetic dyslipidaemia and mixed dyslipidaemia, heart failure,hypercholesterolaemia, cardiovascular disease including atherosclerosis,arteriosclerosis, and hypertriglyceridaemia. As such, the compounds mayalso find favour as therapeutics for coronary artery disease,thrombosis, angina, chronic renal failure, peripheral vascular diseaseand stroke, as well as the cardiovascular indications associated withtype II diabetes mellitus, type I diabetes, insulin resistance,hyperlipidaemia, anorexia nervosa, obesity.

It will be appreciated that references herein to treatment extend toprophylaxis, prevention of recurrence and suppression of symptoms aswell as the treatment of established conditions.

In a further or alternative aspect there is provided a method for thetreatment of a human or animal subject with a condition in whichunder-activation of the HM74A receptor contributes to the condition orin which activation of the receptor will be beneficial, which methodcomprises administering to said human or animal subject an effectiveamount of a compound of Formula (I) or a physiologically acceptable saltor solvate thereof, or a compound of Formula (Ia) or a pharmaceuticallyacceptable salt, solvate or physiologically functional derivativethereof.

More particularly, the present invention provides a method for thetreatment of disorders of lipid metabolism including dislipidaemia orhyperlipoproteinaemia such as diabetic dyslipidaemia and mixeddyslipidaemia, heart failure, hypercholesterolaemia, cardiovasculardisease including atherosclerosis, arteriosclerosis, orhypertriglyceridaemia which method comprises administering to said humanor animal subject an effective amount of a compound of Formula (I) or aphysiologically acceptable salt or solvate thereof or a compound ofFormula (Ia) or a pharmaceutically acceptable salt, solvate orphysiologically functional derivative thereof. The invention alsoprovides methods for the treatment of coronary artery disease,thrombosis, angina, chronic renal failure, peripheral vascular diseaseor stroke, as well as the cardiovascular indications associated withtype II diabetes mellitus, type I diabetes, insulin resistance,hyperlipidaemia, anorexia nervosa, obesity which methods compriseadministering to said human or animal subject an effective amount of acompound of Formula (I) or a physiologically acceptable salt, solvate orderivative thereof or a compound of Formula (Ia) or a pharmaceuticallyacceptable salt, solvate or physiologically functional derivativethereof.

The amount of a HM74A modulator which is required to achieve the desiredbiological effect will, of course, depend on a number of factors, forexample, the mode of administration and the precise clinical conditionof the recipient. In general, the daily dose will be in the range of 0.1mg-1 g/kg, typically 0.1-100 mg/kg. An intravenous dose may, forexample, be in the range of 0.01 mg to 0.1 g/kg, typically 0.01 mg to 10mg/kg, which may conveniently be administered as an infusion of from 0.1μg to 1 mg, per minute. Infusion fluids suitable for this purpose maycontain, for example, from 0.01 μg to 0.1 mg, per millilitre. Unit dosesmay contain, for example, from 0.01 μg to 1 g of a HM74A modulator. Thusampoules for injection may contain, for example, from 0.01 μg to 0.1 gand orally administrable unit dose formulations, such as tablets orcapsules, may contain, for example, from 0.1 mg to 1 g. No toxicologicaleffects are indicated/expected when a compound of the invention isadministered in the above mentioned dosage range.

A compound of the present invention may be employed as the compound perse in the treatment of a the treatment of diseases whereunder-activation of the HM74A receptor contributes to the disease orwhere activation of the receptor will be beneficial, but is preferablypresented with an acceptable carrier in the form of a pharmaceuticalformulation. The carrier must, of course, be acceptable in the sense ofbeing compatible with the other ingredients of the formulation and mustnot be deleterious to the recipient. The carrier may be a solid or aliquid, or both, and is preferably formulated with the HM74A modulatoras a unit-dose formulation, for example, a tablet, which may containfrom 0.05% to 95% by weight of the HM74A modulator.

The formulations include those suitable for oral, rectal, topical,buccal (e.g. sub-lingual) and parenteral (e.g. subcutaneous,intramuscular, intradermal or intravenous) administration.

There is also provided according to the invention a process forpreparation of such a pharmaceutical composition which comprises mixingthe ingredients.

Formulations suitable for oral administration may be presented indiscrete units, such as capsules, cachets, lozenges or tablets, eachcontaining a predetermined amount of a HM74A modulator; as a powder orgranules; as a solution or a suspension in an aqueous or non-aqueousliquid; or as an oil-in-water or water-in-oil emulsion. In general, theformulations are prepared by uniformly and intimately admixing theactive HM74A modulator with a liquid or finely divided solid carrier, orboth, and then, if necessary, shaping the product. For example, a tabletmay be prepared by compressing or moulding a powder or granules of theHM74A modulator optionally with one or more accessory ingredients.Compressed tablets may be prepared by compressing, in a suitablemachine, the compound in a free-flowing form, such as a powder orgranules optionally mixed with a binder, lubricant, inert diluent and/orsurface active/dispersing agent(s). Moulded tablets may be made bymoulding, in a suitable machine, the powdered compound moistened with aninert liquid diluent.

Tablets and capsules for oral administration may contain conventionalexcipients such as binding agents, for example syrup, acacia, gelatin,sorbitol, tragacanth, mucilage of starch or polyvinyl pyrrolidone;fillers, for example, lactose, microcrystalline cellulose, sugar,maize-starch, calcium phosphate or sorbitol; lubricants, for example,magnesium stearate, stearic acid, talc, polyethylene glycol or silica;disintegrants, for example, potato starch, croscarmellose sodium orsodium starch glycollate; or wetting agents such as sodium laurylsulphate. The tablets may be coated according to methods well known inthe art. Oral liquid preparations may be in the form of, for example,aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, ormay be presented as a dry product for constitution with water or othersuitable vehicle before use. Such liquid preparations may containconventional additives such as suspending agents, for example, sorbitolsyrup, methyl cellulose, glucose/sugar syrup, gelatin, hydroxymethylcellulose, carboxymethyl cellulose, aluminium stearate gel orhydrogenated edible fats; emulsifying agents, for example, lecithin,sorbitan mono-oleate or acacia; non-aqueous vehicles (which may includeedible oils), for example almond oil, fractionated coconut oil, oilyesters, propylene glycol or ethyl alcohol; or preservatives, forexample, methyl or propyl p-hydroxybenzoates or sorbic acid. Thepreparations may also contain buffer salts, flavouring, colouring and/orsweetening agents (e.g. mannitol) as appropriate.

Formulations suitable for buccal (sub-lingual) administration includelozenges comprising a HM74A modulator in a flavoured base, usuallysucrose and acacia or tragacanth, and pastilles comprising the HM74Amodulator in an inert base such as gelatin and glycerin or sucrose andacacia.

Formulations of the present invention suitable for parenteraladministration conveniently comprise sterile aqueous preparations of anHM74A modulator, preferably isotonic with the blood of the intendedrecipient. These preparations are preferably administered intravenously,although administration may also be effected by means of subcutaneous,intramuscular, or intradermal injection. Such preparations mayconveniently be prepared by admixing the HM74A modulator with water andrendering the resulting solution sterile and isotonic with the blood.Injectable compositions according to the invention will generallycontain from 0.1 to 5% w/w of the HM74A modulator.

Thus, formulations of the present invention suitable for parenteraladministration comprising a compound according to the invention may beformulated for parenteral administration by bolus injection orcontinuous infusion and may be presented in unit dose form, for instanceas ampoules, vials, small volume infusions or pre-filled syringes, or inmulti-dose containers with an added preservative. The compositions maytake such forms as solutions, suspensions, or emulsions in aqueous ornon-aqueous vehicles, and may contain formulatory agents such asanti-oxidants, buffers, antimicrobial agents and/or toxicity adjustingagents. Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g. sterile, pyrogen-free water,before use. The dry solid presentation may be prepared by filling asterile powder aseptically into individual sterile containers or byfilling a sterile solution aseptically into each container andfreeze-drying.

Formulations suitable for rectal administration are preferably presentedas unit-dose suppositories. These may be prepared by admixing a HM74Amodulator with one or more conventional solid carriers, for example,cocoa butter or glycerides and then shaping the resulting mixture.

Formulations suitable for topical application to the skin preferablytake the form of an ointment, cream, lotion, paste, gel, spray, aerosol,or oil. Carriers which may be used include vaseline, lanolin,polyethylene glycols, alcohols, and combinations of two or more thereof.The HM74A modulator is generally present at a concentration of from 0.1to 15% w/w of the composition, for example, from 0.5 to 2%.

By topical administration as used herein, we include administration byinsufflation and inhalation. Examples of various types of preparationfor topical administration include ointments, creams, lotions, powders,pessaries, sprays, aerosols, capsules or cartridges for use in aninhaler or insufflator or drops (e.g. eye or nose drops).

Ointments and creams may, for example, be formulated with an aqueous oroily base with the addition of suitable thickening and/or gelling agentsand/or solvents. Such bases may thus, for example, include water and/oran oil such as liquid paraffin or a vegetable oil such as arachis oil orcastor oil or a solvent such as a polyethylene glycol. Thickening agentswhich may be used include soft paraffin, aluminium stearate, cetostearylalcohol, polyethylene glycols, microcrystalline wax and beeswax.

Lotions may be formulated with an aqueous or oily base and will ingeneral also contain one or more emulsifying agents, stabilising agents,dispersing agents, suspending agents or thickening agents.

Powders for external application may be formed with the aid of anysuitable powder base, for example, talc, lactose or starch. Drops may beformulated with an aqueous or non-aqueous base also comprising one ormore dispersing agents, solubilising agents or suspending agents.

Spray compositions may be formulated, for example, as aqueous solutionsor suspensions or as aerosols delivered from pressurised packs, with theuse of a suitable propellant, e.g. dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane,1,1,1,2,3,3,3-heptafluoropropane, 1,1,1,2-tetrafluorethane, carbondioxide or other suitable gas.

Capsules and cartridges for use in an inhaler or insufflator, of forexample gelatin, may be formulated containing a powder mix of a compoundof the invention and a suitable powder base such as lactose or starch.

The pharmaceutical compositions according to the invention may also beused in combination with other therapeutic agents, for example incombination with other classes of dyslipidaemic drugs (e.g. statins,fibrates, bile-acid binding resins or nicotinic acid).

The compounds of the instant invention may be used in combination withone or more other therapeutic agents for example in combination withother classes of dyslipidaemic drugs e.g.3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) orfibrates or bile acid binding resins or nicotinic acid. The inventionthus provides, in a further aspect, the use of such a combination in thetreatment of diseases in which under-activation of the HM74A receptorcontributes to the disease or in which activation of the receptor willbe beneficial and the use of a compound of Formula (I) or apharmaceutically acceptable salt, solvate or physiologically functionalderivative thereof, or a compound of Formula (Ia) or a pharmaceuticallyacceptable salt, solvate or physiologically functional derivativethereof, in the manufacture of a medicament for the combination therapyof disorders of lipid metabolism including dislipidaemia orhyperlipoproteinaemia such as diabetic dyslipidaemia and mixeddyslipidaemia, heart failure, hypercholesterolaemia, cardiovasculardisease including atherosclerosis, arteriosclerosis, orhypertriglyceridaemia, coronary artery disease, thrombosis, angina,chronic renal failure, peripheral vascular disease or stroke, as well asthe cardiovascular indications associated with type II diabetesmellitus, type I diabetes, insulin resistance, hyperlipidaemia, anorexianervosa, obesity.

When the compounds of the present invention are used in combination withother therapeutic agents, the compounds may be administered eithersequentially or simultaneously by any convenient route.

The combinations referred to above may conveniently be presented for usein the form of a pharmaceutical formulation and thus pharmaceuticalformulations comprising a combination as defined above optimallytogether with a pharmaceutically acceptable carrier or excipientcomprise a further aspect of the invention. The individual components ofsuch combinations may be administered either sequentially orsimultaneously in separate or combined pharmaceutical formulations.

When combined in the same formulation it will be appreciated that thetwo components must be stable and compatible with each other and theother components of the formulation and may be formulated foradministration. When formulated separately they may be provided in anyconvenient formulation, conveniently in such a manner as are known forsuch compounds in the art.

When in combination with a second therapeutic agent active against thesame disease, the dose of each component may differ from that when thecompound is used alone. Appropriate doses will be readily appreciated bythose skilled in the art.

The invention thus provides, in a further aspect, a combinationcomprising a compound of Formula (I) or a physiologically acceptablesalt or solvate thereof together, or a compound of Formula (Ia) or apharmaceutically acceptable salt, solvate or physiologically functionalderivative thereof, with another therapeutically active agent. Thecombination may conveniently be presented for use in the form of apharmaceutical formulation and thus pharmaceutical formulationscomprising a combination as defined above together with apharmaceutically acceptable carrier thereof represent a further aspectof the invention.

Compounds of Formula (I) and (Ia), and salts and solvates thereof, maybe prepared by various synthetic routes, including the methodologydescribed hereinafter which constitutes a further aspect of theinvention.

Abbreviations

-   -   PyHOTs Pyridinium tosylate    -   CDI Carbonyl diimidazole    -   THF Tetrahydrofuran    -   DCM Dichloromethane    -   TFA Trifluoroacetic Acid

Method A:

A process for preparing compounds of Formula (I)

in which R represents tetrazole, R¹ represents H, Z represents —Y—W—X—,Y represents —(CH₂)_(p)O—, X is absent and W and R² each representphenyl is set out in scheme (a). This method is also suitable forpreparing compounds of Formula (Ia) in which R represents CO₂H, R¹represents C₁-C₅alkoxy, Z represents —Y—W—X—, Y represents —(CH₂)_(p)O—,X is absent and W and R² each represent phenyl:

Accordingly, the present invention provides a process for preparing acompound of Formula (I) comprising coupling of 4′ substitutedanthranilic acid or [2-(1H-tetrazol-5-yl)phenyl]amine with(4-biphenylyloxy)acetic acid using CDI and where desired or necessaryconverting a resultant free acid or base compound of Formula (I) orFormula (Ia) into a physiologically acceptable salt form, or vice versa,or converting one salt form into another physiologically acceptable saltform.

Method B:

A process for preparing compounds of Formula (I)

in which R represents (CH2)_(q)CO₂H, R1 represents H, Z represents—Y—W—X—, Y represents —(CH2)_(p)O—, X is absent and W and R² eachrepresent phenyl is set out in scheme (b):

Accordingly, the present invention provides a process for preparing acompound of Formula (I) comprising coupling of an acid-protected aminewith (4-biphenylyloxy)acetic acid using CDI followed by hydrolysis ofthe protecting group e.g. tert-butyl ester using TFA. and where desiredor necessary converting a resultant free acid or base compound ofFormula (I) into a physiologically acceptable salt form, or vice versa,or converting one salt form into another physiologically acceptable saltform.

The following non-limiting examples illustrate the present invention:

SYNTHETIC EXAMPLES Example 12-{[(4-Biphenylyloxy)acetyl]amino}-4-(methyloxy)benzoic acid

Using method A, a stirred solution of (4-biphenylyloxy)acetic acid (315mg, 1.38 mmol, 1.0 equiv) and CDI (269 mg, 1.66 mmol, 1.2 equiv) inanhydrous THF (10 ml) was left to react at rt. for 60 minutes.2-Amino-4-(methyloxy)benzoic acid (Synth. Commun.; 24, 4; 1994; 533-548)(300 mg, 1.79 mmole, 1.3 equiv) and pyridinium p-toluenesulfonate (833mg, 3.3 mmole, 2.4 equiv) were added and the mixture heated at gentlereflux for 18 hours. The mixture was allowed to cool, filtered and thefiltrate concentrated. CH3CN (15 ml) and THF (3 ml) were added to theresulting residue and then heated to near boiling. After cooling theresulting product was collected by filtration. (321 mg, 65%). δ_(H) H(400 MHz, d6-DMSO) 3.83 (3H, s), 4.78 (2H, s), 6.77 (1H, dd, J=2.4 and9.0 Hz), 7.17 (2H, d, J=8.6 Hz), 7.34 (1H, t, J=7.2 Hz), 7.43 (2H, app.t, J=7.6 Hz), 7.64 (4H, app. t, J=8.8 Hz), 7.98 (1H, d, J=8.8 Hz), 8.36(1H, d, J=2.3 Hz), 12.40 (1H, s), 13.47 (1H, br. s); m/z 378.1 [MH+].

Example 2 2-(4-Biphenylyloxy)-N-[2-(1H-tetrazol-5-yl)phenyl]acetamide

Method A using (4-biphenylyloxy)acetic acid (J. Med. Chem 1976, 19,1079-1088) (128 mg, 0.56 mmol, 1 equiv) and[2-(1H-tetrazol-5-yl)phenyl]amine (118 mg, 0.73 mmol, 1.3 equiv).Crystallisation from acetonitrile afforded the title compound as a whitesolid (93.5 mg, 45%); δ_(H) (400 MHz, DMSO) 4.83 (2H, s), 7.29-7.40 (4H,m), 7.44 (2H, t, J=7.5 Hz), 7.60-7.72 (5H, m), 8.02 (1H, dd, J=8.0 and1.0 Hz), 8.71 (1H, d, J=8.0 Hz), 11.93 (1H, s), one NH not observed upto δ_(H) 13; m/z 372.1 [MH⁺].

Example 3 (2-{[(4-Biphenylyloxy)acetyl]amino}phenyl)acetic acid

a) 1,1-dimethylethyl(2-{[(4-biphenylyloxy)acetyl]amino}phenyl)acetate

Method A using (4-biphenylyloxy)acetic acid (76 mg, 0.33 mmol, 1 equiv)and 1,1-dimethylethyl(2-aminophenyl)acetate (Liebigs Annalen der Chemie,1985, 7, 1398-1412) (90 mg, 0.43 mmol, 1.3 equiv). Purification byBiotage™ chromatography gradient elution with 49-9:1 cyclohexane:ethylacetate afforded the title compound as a colourless gum (73 mg, 55%); δH(400 MHz, CDCl₃) 1.46 (9H, s), 3.44 (2H, s), 4.74 (2H, s), 7.08-7.12(2H, m), 7.16 (1H, app t, J=7.5 Hz), 7.22 (1H, dd, J=7.5 and 1.5 Hz),7.32-7.38 (2H, m), 7.45 (2H, t, J=7.5 Hz), 7.55-7.60 (4H, m), 7.94 (1H,d, J=8.0 Hz), 9.81 (1H, s); m/z 416.2 [M-H]−.

b) (2-{[(4-Biphenylyloxy)acetyl]amino}phenyl)acetic acid

To a solution of1,1-dimethylethyl(2-{[(4-biphenylyloxy)acetyl]amino}phenyl)acetate (30mg, 0.07 mmol) in DCM (0.5 ml) was added TFA (0.5 ml). The mixture wasstirred at room temperature for 3 hours before concentrating in vacuo.The residue was dissolved in DCM (3 ml) and concentrated 3 times toafford the title compound as a pale tan solid (25 mg, 98%); δ_(H) (400MHz, CDCl₃) 3.58 (2H, s), 4.73 (2H, s), 7.01 (2H, dd, J=9.0 and 2.5 Hz),7.20 (1H, app t, J=7.5 Hz), 7.25-7.30 (1H, m), 7.31-7.40 (2H, m),7.41-7.47 (2H, m), 7.47-7.56 (4H, m), 7.79 (1H, d, J=8.0 Hz), 9.34 (1H,s), one exchangeable proton not observed up to δ_(H) 13; m/z 362.1[MH⁺].

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

The application of which this description and claims forms part may beused as a basis for priority in respect of any subsequent application.The claims of such subsequent application may be directed to any featureor combination of features described herein. They may take the form ofproduct, composition, process, or use claims and may include, by way ofexample and without limitation the following claims:

1. A compound selected from: a compound of Formula (I)

and a salt, or hydrate thereof, wherein: R represents (CH₂)_(q)CO₂H or a5 or 6-member aryl, heteroaryl, heterocyclic or alicyclic ring; R¹represents H, halogen, C₁₋₅alkyl, C₁-C₅alkoxy, C₁₋₅haloalkyl, orC₁₋₅haloalkoxy; R² represents a 5 or 6-member aryl, heteroaryl,heterocyclic or alicyclic ring; Z represents —Y—W—X—; W represents a 5or 6-member aryl, heteroaryl, heterocyclic or alicyclic ring; Y is—CH₂O— and X is absent; n represents an integer selected from 2, 3 and4; p represents an integer selected from 0, 1 and 2; q represents aninteger selected from 1, 2, 3 and 4; R³ represents hydrogen or methyl;and R⁴ and R⁵, which may be the same or different, independentlyrepresent C₁-C₃alkyl.
 2. A compound as claimed in claim 1 wherein: Rrepresents CO₂H; and R¹ represents C₁-C₅alkoxy, C₁₋₅haloalkyl, orC₁₋₅haloalkoxy.
 3. A compound according to claim 1 wherein R¹ isselected from C₁-C₃alkoxy, C₁₋₃haloalkyl, and C₁₋₃haloalkoxy.
 4. Acompound according to claim 3 wherein R¹ represents methoxy.
 5. Acompound according to claim 1 wherein R¹ is selected from H, halogen andC₁-C₃alkyl.
 6. A compound according to claim 1 wherein R is selectedfrom pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl,imidazolyl, tetrazolyl, oxazolyl and isoxazolyl.
 7. A compound accordingto claim 1 wherein R represents (CH₂)_(q)CO₂H in which q represents 1.8. A compound according to claim 1 wherein R² is selected fromcyclohexyl, phenyl, pyridinyl, pyrimidinyl, pyridazinyl and isoxazolyl.9. A compound according to claim 1 wherein R² is selected from the groupconsisting of:


10. A compound according to claim 1 wherein R² is selected from thegroup consisting of:


11. A compound according to claim 1 wherein R² represents substitutedphenyl.
 12. A compound according to claim 11 wherein R² representsphenyl substituted with one or two substituents selected from halogenC₁₋₃alkyl, C₁₋₃haloalkyl C₁₋₃alkoxy and C₁₋₃ haloalkoxy.
 13. A compoundaccording to claim 1 wherein W is a 5 or 6 member aryl or heteroarylring.
 14. A compound according to claim 13 wherein W is phenyl.
 15. Acompound according to claim 13 wherein W is a 5 member heteroaryl ring.16. A compound of formula (I) as claimed in claim 1 selected from:2-{[(4-Biphenylyloxy)acetyl]amino}-4-(methyloxy)benzoic acid;2-(4-Biphenylyloxy)-N-[2-(1H-tetrazol-5-yl)phenyl]acetamide;(2-{[(4-Biphenylyloxy)acetyl]amino}phenyl)acetic acid. 17-21. (canceled)22. A method for the treatment of a human or animal subject having acondition characterised by under-activation of the HM74A receptor or inwhich activation of the receptor will be beneficial, which methodcomprises administering to said human or animal subject an effectiveamount of the compound according to claim
 1. 23. A method according toclaim 22 wherein the condition is a disorder of lipid metabolismincluding dislipidaemia or hyperlipoproteinaemia or an inflammatorydisease or condition.
 24. A pharmaceutical formulation comprising thecompound according to claim 1 in admixture with one or morephysiologically acceptable diluents, excipients or carriers.
 25. Acombination for administration together or separately, sequentially orsimultaneously in separate or combined pharmaceutical formulations, saidcombination comprising the compound according to claim 1 together withanother therapeutically active agent.
 26. A pharmaceutical formulationcomprising the compound according to claim 1, plus a further activeingredient selected from the group consisting of a statin, a fibrate, abile-acid binding resin and nicotinic acid and one or morephysiologically acceptable diluents, excipients or carriers.